In article <dmachugh-2410561414320001 at gen153026.gen.tcd.ie> dmachugh at mail.tcd.ie (David MacHugh) writes:
>From: dmachugh at mail.tcd.ie (David MacHugh)
>Subject: Isopropanol & EDTA
>Date: 6 Aug 1997 15:56:08 GMT
>>Hi,
>Does anyone know whether very high concentrations of EDTA (~0.25M) will
>affect DNA precipitations using isopropanol? I have noticed that the
>aqeuous material seems to separate out as a layer below the isopropanol.
>I haven't tried to recover the DNA yet, but I reckon it won't work.
>Any comments..
>Thanks in advance.
>David MacHugh.
>Genetics Department,
>TCD.
>Ireland.
Yeah, any high salt will result in an aqueues layer forming with isopropanol
(the concentration of IPA to obtain this will depend upon the net saly
concentration). However, you can still get your dna recovered by diluting the
aqueus layer with 0.3M NaOAc to reduce the salt content and then adding IPA to
50% final.
However, if youve already ppt the EDTA it is unlikely to be washed away in 70%
EtOH..in which case ppt, resolvate in excess TE or dH2O and reppt!
Good luck
Rob